For live imaging, transgenic embryos were mounted in a drop of 1% low melting agarose in filtered artificial seawater on the surface of a 35-mm glass bottom dish (Ibidi μ-Dish) with the eye facing the glass surface.
Confocal images were obtained on a Zeiss LSM780 laser scanning confocal microscope. For live imaging, we used a Zeiss C-Apochromat 40x NA 1.2 water immersion objective, while immunostained samples were imaged using Zeiss Plan-Apochromat 40x NA 1.3 oil, Plan-Apochromat 63x NA 1.4 oil and LD LCI Plan-Apochromat 25x NA 0.8 objectives.
Image data were handled using Fiji [92 (link)]; the Enhance Local Contrast (CLAHE) plugin was used to enhance contrast. The confocal images shown in Figs.4 b, c, 5 a, c, and 6 a, d were deconvolved using the Fiji plugin DeconvolutionLab2 [93 (link)]; point spread function was calculated theoretically by the PSF generator plugin [94 (link)]. The 3D rendering shown in Fig. 6 d was obtained using the ImageJ 3D viewer plugin.
A confocal image stack of an adult brain, immunostained for acetylated tubulin, was used for the 3D reconstruction of the Parhyale optic lobes shown in Fig.5 g. Neuropils were labelled manually using the TrakEM2 Fiji plugin [95 (link)].
Confocal images were obtained on a Zeiss LSM780 laser scanning confocal microscope. For live imaging, we used a Zeiss C-Apochromat 40x NA 1.2 water immersion objective, while immunostained samples were imaged using Zeiss Plan-Apochromat 40x NA 1.3 oil, Plan-Apochromat 63x NA 1.4 oil and LD LCI Plan-Apochromat 25x NA 0.8 objectives.
Image data were handled using Fiji [92 (link)]; the Enhance Local Contrast (CLAHE) plugin was used to enhance contrast. The confocal images shown in Figs.
A confocal image stack of an adult brain, immunostained for acetylated tubulin, was used for the 3D reconstruction of the Parhyale optic lobes shown in Fig.
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